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Squidly: Enzyme Catalytic Residue Prediction Harnessing a Biology-Informed Contrastive Learning Framework

Rieger, W. J.; Boden, M.; Arnold, F. H.; Mora, A.

2025-06-20 bioinformatics
10.1101/2025.06.13.659624 bioRxiv
Show abstract

Enzymes present a sustainable alternative to traditional chemical industries, drug synthesis, and bioremediation applications. Because catalytic residues are the key amino acids that drive enzyme function, their accurate prediction facilitates enzyme function prediction. Sequence similarity-based approaches such as BLAST are fast but require previously annotated homologs. Machine learning approaches aim to overcome this limitation; however, current gold-standard machine learning (ML)-based methods require high-quality 3D structures limiting their application to large datasets. To address these challenges, we developed Squidly, a sequence-only tool that leverages contrastive representation learning with a biology-informed, rationally designed pairing scheme to distinguish catalytic from non-catalytic residues using per-token Protein Language Model embeddings. Squidly surpasses state-of-the-art ML annotation methods in catalytic residue prediction while remaining sufficiently fast to enable wide-scale screening of databases. We ensemble Squidly with BLAST to provide an efficient tool that annotates catalytic residues with high precision and recall for both in- and out-of-distribution sequences.

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